SUMMARY OF WORK: Understanding the relationships between structure and function for a DNA enzyme holds the promise of allowing us to develop specific inhibitors and other modulators of the enzymes activity through rational design approaches. Although this idea is now fundamental in the field of structural biology, we are still in the earliest stages of bringing the approach to its full potential.Our studies of mammalian DNA polymerase beta have pioneered the use of a coordinated approach of structural studies (x-ray crystallography, NMR, and spectroscopy), biochemical studies and mammalian genetic studies. This approach has allowed us to establish the cellular role of DNA polymerase beta in mammalian base excision repair. And, the approach has allowed us to establish a solid framework for future studies of the role of individual amino acid residues in this enzyme in such important endpoints as the rate of DNA repair, coordination of DNA synthesis with DNA ligation, coordination of deoxyribose phosphate removal (lyase activity) with DNA synthesis, the fidelity of DNA synthesis, the fidelity of overall DNA base excision repair, and DNA lesion bypass. Rational drug design targeting these endpoints will allow us to strategically regulate base excision repair with DNA polymerase beta specific drugs.Detailed structure-function relationship studies of other base excision repair enzymes, such as XRCC1, DNA ligases I and III, AP endonuclease, and the various DNA glycosylases, will be undertaken in the future. Development of specific inhibitors or other modulators for these enzymes will allow us to strategically deregulate base excision repair in cells. This could have implications for chemotherapy and for understanding the role of DNA repair in preventing disease especially after exposure to environmental toxins. - base excision repair, DNA polymerase beta, structure-function, structural biology, fidelity, AP endonuclease, DNA synthesis, lyase, site-directed mutagenesis